Indirect Shoot Regeneration Using 2,4-D Induces Somaclonal Variations in Cinchona officinalis

Abstract

Cinchona officinalis is an important species from the Andean cloud forest that has a low regeneration rate in natural populations. In vitro regeneration of C. officinalis has been successfully established but somaclonal variation was not evaluated. The regeneration pathway and the number of subcultures on somaclonal variation were evaluated using six ISSR primers that amplified 58 loci of Inter Simple Sequence Repeats (ISSR). A dendrogram based on Jaccard´s genetic distance between the subcultures and the donor plant was produced. The results show that indirect shoot regeneration induces somaclonal variation, in the presence 2,4-Dichlorophenoxyacetic acid (2,4-D) in combination with kinetin and 6-Benzylaminopurine (BAP). In combination with 1-Naphthaleneacetic acid (NAA) or with Indole-3-butyric acid (IBA), BAP produces genetically stable explants. The highest proliferation rate was achieved using BAP and IBA. The present research study suggests avoiding the use of 2,4-D when C. officinalis is propagated for reintroduction and restoration projects.

Efficient shoot regeneration of medicinal plant Haplophyllum tuberculatum by direct and indirect organogenesis and genetic fidelity assessment using Inter Simple Sequence Repeats markers

“…Although a high concentration of 2,4-D resulted in profuse callus growth, a high concentration of 2,4-D can decrease regeneration frequency, and calluses tend to remain in the callus proliferation stage. And there is an increased risk of the development of somaclonal variations under elevated levels of 2,4-D in growth media (Armijos-González et al, 2021), which was also observed during our experiments after assessing regenerated plants with ISSR markers. Therefore 2,4-D used for H. tuberculatum calli production is probably one of the major factors that affect the genetic fidelity of regenerated shoots.…”

“…Therefore 2,4-D used for H. tuberculatum calli production is probably one of the major factors that affect the genetic fidelity of regenerated shoots. Thus the observed somaclonal variations in regenerated shoots may be due to the heterogeneity of regenerated callus cells and seeds ( Armijos-González et al, 2021 ). In other Haplophyllum species such as Haplophyllum virgatum var.…”

“…The plant species ( H. tuberculatum ) was identified accurately using guidelines from the text book “An Illustrated Checklist of the Flora of Qatar” ( Norton et al, 2009 ) and verified by a plant taxonomist. All the explants for the rest of the experiments were obtained from the in vitro raised plantlets ( Armijos-González et al, 2021 ).…”